Development of transgenic rabbits as models for atherosclerotic research

开发转基因兔作为动脉粥样硬化研究模型

• 批准号: 08557017
• 负责人: WATANABE Teruo
• 金额: $ 9.28万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08557017/
• 关键词: Transgenic Animals; Lipoproteins; Hypercholesterolemia; Rabbit; Apoproteins; Gene expression; Cloning; トランスジェニックウサギ; レムナント; 冠状動脈硬化; 危険因子; リボ蛋白; リボ蛋白(a)

For the study of atheroslerosis, we have successfully generated transgenic rabbits expressing either human apolipoprotein(a) [Lp(a)] or lipoprotein lipase (LPL) at the end of 1998.Apo(a) transgenic rabbits :Totally, 93 pups were obtained after transplantation into foster mothers. 11 rabbits were found to have transgene integrated into their genome. 3 transgenic rabbits soon after birth and 5 transgenic rabbits show no detectable expression of apo(a) in the plasma. 3 transgenic rabbit lines have been established ; A01 line has 1.8mg/dl, A46 line has 3.4 mg/dI, and A47 line has 4.5 mg/dl of apo(a) in the plasma. A10 line has been bred into F1 and F2 and the following results were obtained. Human apo(a) mRNA expression was mainly found in the liver and kidney. Plasma apo(a), in transgenic rabbits showed a similar mobility to human Lp(a) at pre-beta position. In the plasma of transgenic rabbits, unlike the plasma of transgenic mice, about 80% of the apo(a) was covalently associated with rabbit apo-B and was contained in the fractions with density 1.02-1.10 g/ml, indicating the formation of Lp(a). These results suggest that transgenic rabbits expressing human apo(a) exhibit efficient assembly of Lp(a) and can be used as an animal model for the study of human Lp(a). Now, we have started to breed more F1 for the study of atherosclerosis by feeding cholesterol-rich diet.LPL transgenic rabbits :Until now, we have produced 3 LPL transgenic rabbits under the control of chicken b-actin promoter and 4 LPL transgenic rabbits under the control of scavenger receptor promoter. Characterization of these transgenic rabbits are currently underway.

为研究动脉粥样硬化，我们于1998年底成功地培育了表达人载脂蛋白（a）[Lp（a）]或脂蛋白脂酶（LPL）的转基因兔。11只家兔的基因组中整合了转基因。3只出生后不久的转基因兔和5只转基因兔血浆中未检测到apo（a）的表达。建立了3个转基因兔系，A01系血浆apo（a）浓度为1.8mg/dl，A46系为3.4mg/dl，A47系为4.5mg/dl。用A10系配组F1和F2，结果如下：人apo（a）mRNA主要表达于肝和肾。转基因兔血浆载脂蛋白（a）在前β位置显示与人Lp（a）相似的迁移率。在转基因兔血浆中，与转基因小鼠血浆不同，约80%的apo（a）与兔apo-B共价结合，并包含在密度为1.02-1.10 μ g/ml的组分中，表明形成了Lp（a）。这些结果表明，表达人载脂蛋白（a）的转基因兔表现出高效的Lp（a）组装，可以用作研究人Lp（a）的动物模型。目前，我们已经通过饲喂高胆固醇饲料，开始培育更多的F1代，用于动脉粥样硬化的研究。LPL转基因兔：到目前为止，我们已经获得了3只鸡β-肌动蛋白启动子控制的LPL转基因兔和4只清道夫受体启动子控制的LPL转基因兔。这些转基因兔子的特征目前正在进行中。

项目成果

Matsumoto, S., Katoh, M., Saito, S., Watanabe, T., Masuho, Y.: "Identification of soluble type of membrane-type matrix metalloproteinase-3 formed by alternative spliced mRNA,Biochem." Biochem Biophys Acta. 1354. 159-170 (1997)

Matsumoto, S.、Katoh, M.、Saito, S.、Watanabe, T.、Masuho, Y.：“鉴定由可变剪接 mRNA 形成的可溶型膜型基质金属蛋白酶 3，Biochem。”

范,江霖、渡辺照男: "トランスジェニックウサギ。動脈硬化+高脂血症ストラテジー。" 細胞工学別冊 山田信博、佐藤靖史監修 秀潤社. 447-449 (1996)

Fan、Jiang Lin、Teruo Watanabe：“转基因兔。动脉硬化+高脂血症策略。”，Nobuhiro Yamada 和 Yasushi Sato 编辑，Shujunsha 447-449 (1996)。

范 江霖、渡辺照男: "トランスジェニックウサギ、動脈硬化+高脂血症研究ストラテジー" 細胞工学別冊 山田信博、佐藤靖史監修. 秀潤社. 447-449 (1996)

范江林，渡边辉夫：“转基因兔，动脉硬化+高脂血症研究策略”细胞工程特刊，山田信宏和佐藤康史监督447-449（1996）。

Araki M,Fan J,Challah M,Bensadoun A,Yamada N,Kakuta H,Shikama H and Watanabe T: "Transgenic rabbits expressing human lipoprotein lipase." Cytotechnology. (in press).

Araki M、Fan J、Challah M、Bensadoun A、Yamada N、Kakuta H、Shikama H 和 Watanabe T：“表达人脂蛋白脂肪酶的转基因兔子。”

Fan J.Challah M and Watanabe T: "Transgenic rabbit models for biomedical research. Current status, basic methods and future perspective." Pathol Internat. (in press).

Fan J.Challah M 和 Watanabe T：“用于生物医学研究的转基因兔模型。现状、基本方法和未来展望。”